Construction of a full length cDNA library from the skeletal muscle of Larimichthys crocea and its EST analysis

A full-length cDNA library was constructed for the skeletal muscle of Larimichthys crocea using the Creator Smart cDNA Library Construction Kit.Library quality analysis showed that the library reached 1.68×106 cfu/mm3 in capacity with a recombinant efficiency as high as 96%.PCR results showed that the average size of inserts was larger than 1 000 bp.512 clones were randomly selected and sequenced.Of these 430 were expressed sequence tags(ESTs) longer than 100 bp.From these ESTs,230 non-repetitive sequences were acquired,including 58 contigs and 172 singletons after initial assembly and with a redundancy rate of 46.51%.From this retrieval 35 ESTs seemed to be novel genes(E-value≤1.00×10-10) as no clearly homologous known sequence was found in GenBank databases;195 ESTs were found to have considerable homology to reported genes,of which energy-related genes had the highest abundance,accounting for 20.00%,while protein synthesis,cytoskeleton and signal transduction genes made up 13.48%,12.17% and 10.00%,respectively.These included the high-mobility group protein B1,leptin receptor and β1-heat shock protein.The results provide a foundation for further screening and cloning of specific expression genes in the skeletal muscle of Larimichthys crocea.